Method for storage and stabilization of a target substance

Patent metadata

Field	Value
Publication number	US-9797871-B2
Application number	US-201113813800-A
Country	US
Kind code	B2
Filing date	Apr 28, 2011
Priority date	Aug 5, 2010
Publication date	Oct 24, 2017
Grant date	Oct 24, 2017

Abstract

The invention relates to a system and method for the stable storage of sensitive biological or chemical target substance, in a bound form on certain capture media. The method comprised providing a sample containing the target substance in a suitable buffer; combining the sample with a capture media to effect reversible binding of the target substance to the capture media; and storing the capture media with the target substance at between about −20 and 20° C.; and recovering the target substance from the capture media. The target substance recovered maintains the desired activity. Also provides are methods for reducing aggregates in the sensitive biological or chemical target substance.

First claim

What is claimed is:

1. A method for reducing aggregates in a target substance during a wet storage, the method comprising:
(a) providing a sample containing said target substance in a suitable buffer;
(b) combining said sample with a capture media to effect reversible binding of the target substance to the capture media with the target substance in a hydrated state;
(c) storing said capture media with the target substance reversibly bound thereto in a hydrated state, at between about −20 and 20° C., for at least an amount of time which allows a significant amount of aggregates of the target substance to separate into monomeric form; and
(d) recovering said target substance from said capture media,
wherein the target substance recovered contains a significantly less amount of aggregated form of the target substance compared to the target substance before the storage, is not denatured throughout the method, and maintains a desired activity, and
wherein the capture media is a protein A based affinity media or a multimodal strong anion exchange media that comprises a ligand comprising N-Benzyl-N-methyl ethanol amine.

2. The method of claim 1 , wherein the target substance is monoclonal or polyclonal antibody.

3. The method of claim 1 , wherein the target substance at the combining step is at a concentration of about 1 to 300 g/L (mg/mL) volume.

4. The method of claim 1 , wherein said sample is selected from the group of cell culture supernatant, cell culture lysate, bacterial culture supernatant, bacterial culture lysate, blood or other body fluids.

5. The method of claim 1 , wherein combining step is performed in a container selected from column, bag, tank, cassette, or other housing and the container is capable of supporting the binding between the target substance and the capture media, as well as the storage step.

6. The method of claim 1 , wherein the capture media with the target substance reversibly bound thereto is stored for less than 2 weeks.

7. The method of claim 1 , wherein the capture media with the target substance reversibly bound thereto is stored for about 2-4 weeks.

8. The method of claim 1 , wherein the recovering step includes eluting the target substance from the capture media and collecting the eluent containing the target substance.

9. The method of claim 1 , wherein the capture media comprises highly cross-linked agarose.

10. The method of claim 1 , wherein the recovery rate is at least 75%.

11. The method of claim 1 , wherein the recovery rate is at least 50%.

12. The method of claim 1 , wherein the combining step is performed in a container selected from column, bag, tank, cassette, or other housing and the container is capable of supporting the binding between the target substance and the capture media as well as the storage step.

13. The method of claim 12 , wherein said container is transportable and/or can withstand freezing.

14. The method of claim 1 , wherein the capture media with the target substance reversibly bound thereto is stored at about −20° C., 4° C., or 20° C.

15. The method of claim 1 , wherein the capture media with the target substance reversibly bound thereto may be stored at temperatures up to 40° C.

16. The method of claim 1 , wherein the capture media with the target substance reversibly bound thereto is stored for more than 4 weeks.

17. A method for reducing aggregates in a target substance during a wet storage, the method comprising:
(a) providing a sample containing said target substance in a suitable buffer;
(b) combining said sample with a capture media to effect reversible binding of the target substance to the capture media with the target substance in a hydrated state;
(c) storing said capture media with the target substance reversibly bound thereto in a hydrated state, at between about −20 and 20° C., for at least a day; and
(d) recovering said target substance from said capture media,
wherein the target substance recovered contains a significantly less amount of aggregated form of the target substance compared to the target substance before the storage, is not denatured throughout the method, and maintains a desired activity, and
wherein the capture media is affinity media.

18. The method of claim 17 , wherein the target substance is monoclonal or polyclonal antibody.

19. The method of claim 17 , wherein the target substance at the combining step is at a concentration of about 1 to 300 g/L (mg/mL) volume.

20. The method of claim 17 , wherein said sample is selected from the group of cell culture supernatant, cell culture lysate, bacterial culture supernatant, bacterial culture lysate, blood or other body fluids.

21. The method of claim 17 , wherein combining step is performed in a container selected from column, bag, tank, cassette, or other housing and the container is capable of supporting the binding between the target substance and the capture media, as well as the storage step.

22. The method of claim 17 , wherein the capture media with the target substance reversibly bound thereto is stored for less than 2 weeks.

23. The method of claim 17 , wherein the capture media with the target substance reversibly bound thereto is stored for more than 2 weeks.

24. The method of claim 17 , wherein the recovering step includes eluting the target substance from the capture media and collecting the eluent containing the target substance.

25. The method of claim 17 , wherein the capture media comprises highly cross-linked agarose.

26. The method of claim 17 , wherein the recovery rate is at least 75%.

27. The method of claim 17 , wherein the recovery rate is at least 50%.

28. The method of claim 17 , wherein the target substance is selected from proteins, peptides, oligopeptides, oligonucleotides, RNA, DNA, protein vaccine, virus vaccine, and other sensitive therapeutic substance.

29. A method for reducing aggregates in a target substance during a wet storage, the method comprising:
(a) providing a sample containing said target substance in a suitable buffer;
(b) combining said sample with a capture media to effect reversible binding of the target substance to the capture media with the target substance in a hydrated state;
(c) storing said capture media with the target substance reversibly bound thereto in a hydrated state, at between about −20 and 20° C., for at least a day; and
(d) recovering said target substance from said capture media,
wherein the target substance recovered contains a significantly less amount of aggregated form of the target substance compared to the target substance stored in solution, is not denatured throughout the method, and maintains a desired activity.

30. The method of claim 29 , wherein target substance is bound to the capture media through ion-exchange interaction, cation-pi interaction, pi-pi interaction, hydrogen bonding interaction, metal ion affinity, hydrophobic interaction, boronate glycomolecule affinity, or combinations of such interactions via mixed mode, hydroxyapatite, blue dye ligand, or affinity interactions including lectin and protein ligand based affinities.

Assignees

• Van Alstine James
• Ohman Johan
• Busson Philippe
• Palmgren Ronnie
• Allmer Klas
• Daicic John
• Ge Healthcare Bio Sciences Ab

Inventors

• Van Alstine James
• Ohman Johan
• Busson Philippe
• Palmgren Ronnie
• Allmer Klas
• Daicic John

Classifications

• C07K1/22

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• G01N30/56 Primary

  Packing methods or coating methods · CPC title

• B01D15/206

  Packing or coating · CPC title

• G01N30/6069

  with compartments or bed substructure · CPC title

• G01N2030/562

  packing · CPC title